US8865155B2 - Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom - Google Patents
Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom Download PDFInfo
- Publication number
- US8865155B2 US8865155B2 US13/629,398 US201213629398A US8865155B2 US 8865155 B2 US8865155 B2 US 8865155B2 US 201213629398 A US201213629398 A US 201213629398A US 8865155 B2 US8865155 B2 US 8865155B2
- Authority
- US
- United States
- Prior art keywords
- milk
- nrrl
- fermented
- lactis
- strains
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active - Reinstated, expires
Links
- 241000194035 Lactococcus lactis Species 0.000 title claims abstract description 118
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 63
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 40
- 235000014897 Streptococcus lactis Nutrition 0.000 title claims abstract description 18
- 230000000975 bioactive effect Effects 0.000 title abstract description 19
- 238000004519 manufacturing process Methods 0.000 title abstract description 10
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 230000001580 bacterial effect Effects 0.000 title abstract description 5
- 230000000694 effects Effects 0.000 title description 21
- 230000003276 anti-hypertensive effect Effects 0.000 title description 15
- 241000124008 Mammalia Species 0.000 title description 4
- 235000016709 nutrition Nutrition 0.000 title description 2
- 230000001225 therapeutic effect Effects 0.000 title description 2
- 230000003627 anti-cholesterol Effects 0.000 title 1
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 21
- 230000036772 blood pressure Effects 0.000 claims abstract description 14
- 239000000203 mixture Substances 0.000 claims abstract description 5
- 239000008267 milk Substances 0.000 claims description 81
- 235000013336 milk Nutrition 0.000 claims description 78
- 210000004080 milk Anatomy 0.000 claims description 78
- 235000015140 cultured milk Nutrition 0.000 claims description 45
- 238000000034 method Methods 0.000 claims description 15
- 206010020772 Hypertension Diseases 0.000 claims description 11
- 239000007858 starting material Substances 0.000 claims description 9
- 230000001631 hypertensive effect Effects 0.000 claims description 6
- 108010007622 LDL Lipoproteins Proteins 0.000 claims description 5
- 102000007330 LDL Lipoproteins Human genes 0.000 claims description 5
- 238000000855 fermentation Methods 0.000 claims description 5
- 230000004151 fermentation Effects 0.000 claims description 5
- 241000194036 Lactococcus Species 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 claims description 3
- 229920001184 polypeptide Polymers 0.000 claims 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 abstract description 20
- 235000013305 food Nutrition 0.000 abstract description 12
- 108010028554 LDL Cholesterol Proteins 0.000 abstract description 9
- 235000012000 cholesterol Nutrition 0.000 abstract description 9
- 230000036996 cardiovascular health Effects 0.000 abstract description 6
- 235000013376 functional food Nutrition 0.000 abstract description 6
- 230000003078 antioxidant effect Effects 0.000 abstract description 5
- 235000015872 dietary supplement Nutrition 0.000 abstract description 3
- 239000000825 pharmaceutical preparation Substances 0.000 abstract description 3
- 238000008214 LDL Cholesterol Methods 0.000 abstract description 2
- 238000009472 formulation Methods 0.000 abstract description 2
- 238000011699 spontaneously hypertensive rat Methods 0.000 description 48
- 230000035488 systolic blood pressure Effects 0.000 description 24
- 230000037396 body weight Effects 0.000 description 23
- 108010046377 Whey Proteins Proteins 0.000 description 22
- 102000007544 Whey Proteins Human genes 0.000 description 22
- 239000005862 Whey Substances 0.000 description 21
- FAKRSMQSSFJEIM-RQJHMYQMSA-N captopril Chemical compound SC[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O FAKRSMQSSFJEIM-RQJHMYQMSA-N 0.000 description 21
- 229960000830 captopril Drugs 0.000 description 21
- 235000013365 dairy product Nutrition 0.000 description 20
- 230000035487 diastolic blood pressure Effects 0.000 description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- 235000018102 proteins Nutrition 0.000 description 18
- 238000011282 treatment Methods 0.000 description 17
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 12
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 12
- 239000008213 purified water Substances 0.000 description 12
- 241001465754 Metazoa Species 0.000 description 11
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 10
- 101710138623 Kappa-casein Proteins 0.000 description 10
- 239000013642 negative control Substances 0.000 description 10
- 239000013641 positive control Substances 0.000 description 10
- 230000009467 reduction Effects 0.000 description 10
- 241000700159 Rattus Species 0.000 description 9
- QIAFMBKCNZACKA-UHFFFAOYSA-N N-benzoylglycine Chemical compound OC(=O)CNC(=O)C1=CC=CC=C1 QIAFMBKCNZACKA-UHFFFAOYSA-N 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 238000010586 diagram Methods 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 7
- 240000002605 Lactobacillus helveticus Species 0.000 description 7
- 235000013967 Lactobacillus helveticus Nutrition 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 229940054346 lactobacillus helveticus Drugs 0.000 description 7
- 238000005259 measurement Methods 0.000 description 7
- 239000002904 solvent Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 230000004872 arterial blood pressure Effects 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 238000004007 reversed phase HPLC Methods 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 108010023302 HDL Cholesterol Proteins 0.000 description 5
- 102000014171 Milk Proteins Human genes 0.000 description 5
- 108010011756 Milk Proteins Proteins 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 235000021239 milk protein Nutrition 0.000 description 5
- ZZKNRXZVGOYGJT-VKHMYHEASA-N (2s)-2-[(2-phosphonoacetyl)amino]butanedioic acid Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)CP(O)(O)=O ZZKNRXZVGOYGJT-VKHMYHEASA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 230000000055 hyoplipidemic effect Effects 0.000 description 4
- 239000004310 lactic acid Substances 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000001603 reducing effect Effects 0.000 description 4
- 150000003626 triacylglycerols Chemical class 0.000 description 4
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 235000006708 antioxidants Nutrition 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 208000029078 coronary artery disease Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 235000013861 fat-free Nutrition 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 239000008101 lactose Substances 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 230000002797 proteolythic effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- MRBKEAMVRSLQPH-UHFFFAOYSA-N 3-tert-butyl-4-hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1 MRBKEAMVRSLQPH-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- NLZUEZXRPGMBCV-UHFFFAOYSA-N Butylhydroxytoluene Chemical compound CC1=CC(C(C)(C)C)=C(O)C(C(C)(C)C)=C1 NLZUEZXRPGMBCV-UHFFFAOYSA-N 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 241000194032 Enterococcus faecalis Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000235070 Saccharomyces Species 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- DOFAQXCYFQKSHT-SRVKXCTJSA-N Val-Pro-Pro Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DOFAQXCYFQKSHT-SRVKXCTJSA-N 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 239000002220 antihypertensive agent Substances 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- XOEMATDHVZOBSG-UHFFFAOYSA-N azafenidin Chemical compound C1=C(OCC#C)C(Cl)=CC(Cl)=C1N1C(=O)N2CCCCC2=N1 XOEMATDHVZOBSG-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000002060 circadian Effects 0.000 description 2
- 235000014048 cultured milk product Nutrition 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N formic acid Substances OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 230000007407 health benefit Effects 0.000 description 2
- 230000001077 hypotensive effect Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 235000020191 long-life milk Nutrition 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 230000010349 pulsation Effects 0.000 description 2
- 235000000891 standard diet Nutrition 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000004885 tandem mass spectrometry Methods 0.000 description 2
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 2
- 238000010792 warming Methods 0.000 description 2
- AAXWBCKQYLBQKY-IRXDYDNUSA-N (2s)-2-[[(2s)-2-[(2-benzamidoacetyl)amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-4-methylpentanoic acid Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CNC(=O)C=1C=CC=CC=1)C1=CN=CN1 AAXWBCKQYLBQKY-IRXDYDNUSA-N 0.000 description 1
- CUKWUWBLQQDQAC-VEQWQPCFSA-N (3s)-3-amino-4-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s,3s)-1-[[(2s)-1-[(2s)-2-[[(1s)-1-carboxyethyl]carbamoyl]pyrrolidin-1-yl]-3-(1h-imidazol-5-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-methyl-1-ox Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 CUKWUWBLQQDQAC-VEQWQPCFSA-N 0.000 description 1
- 239000005541 ACE inhibitor Substances 0.000 description 1
- 230000010641 Acidifying Activity Effects 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 102400000344 Angiotensin-1 Human genes 0.000 description 1
- 101800000734 Angiotensin-1 Proteins 0.000 description 1
- 102400000345 Angiotensin-2 Human genes 0.000 description 1
- 101800000733 Angiotensin-2 Proteins 0.000 description 1
- 206010005746 Blood pressure fluctuation Diseases 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-N 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 239000001968 M17 agar Substances 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241000364057 Peoria Species 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- NGFMICBWJRZIBI-JZRPKSSGSA-N Salicin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O1)c1c(CO)cccc1 NGFMICBWJRZIBI-JZRPKSSGSA-N 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- GLEVLJDDWXEYCO-UHFFFAOYSA-N Trolox Chemical compound O1C(C)(C(O)=O)CCC2=C1C(C)=C(C)C(O)=C2C GLEVLJDDWXEYCO-UHFFFAOYSA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 101150088616 acma gene Proteins 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- NGFMICBWJRZIBI-UHFFFAOYSA-N alpha-salicin Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UHFFFAOYSA-N 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- ORWYRWWVDCYOMK-HBZPZAIKSA-N angiotensin I Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 ORWYRWWVDCYOMK-HBZPZAIKSA-N 0.000 description 1
- 229950006323 angiotensin ii Drugs 0.000 description 1
- 229940044094 angiotensin-converting-enzyme inhibitor Drugs 0.000 description 1
- 150000001491 aromatic compounds Chemical class 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000004531 blood pressure lowering effect Effects 0.000 description 1
- 238000009530 blood pressure measurement Methods 0.000 description 1
- 238000012754 cardiac puncture Methods 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000003205 diastolic effect Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000000132 electrospray ionisation Methods 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 210000000750 endocrine system Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 108010016268 hippuryl-histidyl-leucine Proteins 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 208000021822 hypotensive Diseases 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 1
- 108010022197 lipoprotein cholesterol Proteins 0.000 description 1
- 238000001972 liquid chromatography-electrospray ionisation mass spectrometry Methods 0.000 description 1
- 230000009063 long-term regulation Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000009928 pasteurization Methods 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 235000020185 raw untreated milk Nutrition 0.000 description 1
- 239000000837 restrainer Substances 0.000 description 1
- NGFMICBWJRZIBI-UJPOAAIJSA-N salicin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UJPOAAIJSA-N 0.000 description 1
- 229940120668 salicin Drugs 0.000 description 1
- 230000009291 secondary effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 235000021262 sour milk Nutrition 0.000 description 1
- 235000011497 sour milk drink Nutrition 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- YMBCJWGVCUEGHA-UHFFFAOYSA-M tetraethylammonium chloride Chemical compound [Cl-].CC[N+](CC)(CC)CC YMBCJWGVCUEGHA-UHFFFAOYSA-M 0.000 description 1
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 239000005526 vasoconstrictor agent Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 235000021119 whey protein Nutrition 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/10—Peptides having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/231—Lactis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Definitions
- the invention relates to Lactococcus lactis strains for the production of bioactive peptides. More particularly, the invention relates to Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having antihypertensive and cholesterol-lowering effects in mammals and related nutritional and therapeutic products.
- Coronary heart disease which is considered the most common and serious form of cardiovascular disease, is the first cause of death in developed industrialized countries.
- Hypertension and elevated blood cholesterol levels, particularly high low density-density lipoprotein cholesterol (LDL-C) are two of the major modified risk factors for the development of CHD (Department of Health and Human Services, 2000).
- ACE angiotensin-converting enzyme
- LAB growth in milk is dependent on the specific proteolytic systems for the generation of free peptides as a source of nitrogen (Hugenholtz, 2008, Int. Dairy J. 18, 466-475). Indeed, several ACEI peptides and/or with antihypertensive activity derived from milk proteins by the action of Lactobacillus helveticus and Saccharomyces cerevisae (Nakamura et al., 1995, J. Dairy Sci. 78:777-783; Nakamura et al., 1995, J. Dairy Sci. 78:1253-1257) or Lactobacillus helveticus (Sipola et al, 2002, J. Dairy Res.
- Calpis sour milk drink Calpis Co., Japan
- Evolus Valio, Finland
- Calpis sour milk is claimed as suitable for those with mild hypertension and is fermented with Lactobacillus helveticus and Saccharomyces cervesiae and Evolus which is claimed as the first European functional food to help lower blood pressure, also fermented with Lactobacillus helveticus .
- Both fermented milk products contain bioactive peptides responsible for the ACE-inhibition and presented antihypertensive effects in hypertensive rats.
- Lactococcus lactis strains are able to improve the organoleptic characteristics of dairy products since they are responsible for the formation of aromatic compounds (Ayad 2009, Food Microbiol 26, 533-541). Previous studies in our laboratory showed that specific L. lactis strains isolated from native ecosystems were able to produce remarkable aroma profiles in fermented milk (Gutierrez-Méndez et al., 2008, J. Dairy Sci. 91, 49-579). Furthermore, it was reported that a wild L. lactis strain presented ACEI peptides in Mexican Fresco cheese (Torres-Llanez, et al., 2011, J. Dairy Sci., 94: 3794-3800). Also, specific wild L.
- lactis strains were explored for their ability to produce ACEI activity in fermented milk (Rodr ⁇ guez-Figueroa et al., 2010, J. Dairy Sci., 93: 5032-5038; Otte et al., 2011, Int. Dairy J., 21: 229-238).
- fermented milk products produced by L. lactis strains were not tested in vivo to show any health benefits.
- Lactococcus lactis strains NRRL B-50571 and NRRL B-50572 have the capacity to produce bioactive peptides that have remarkable capacity to generate an antihypertensive effect in mammals.
- Such hypotensive peptides do not change arterial blood pressure in subjects with normotensive arterial blood pressure; only hypertensive subjects experiment a reduction in arterial blood pressure.
- the bioactive peptides are a viable option to reduce arterial blood pressure without the secondary effects commonly produced by synthetic drugs.
- bioactive peptides improve cardiovascular health by lowering bad (LDL) cholesterol and present antioxidant properties. Therefore, the mentioned lactic acid bacteria and the bioactive peptides included in this invention may be used in pharmaceutical preparations as well as in food products, such as functional foods.
- One or more embodiments include the generation of bioactive peptides by the action of novel Lactococcus lactis NRRL B-50571 or NRRL B-50572 on a substrate comprising one or more proteins or its fragments, which contain specific amino acid sequences. These peptides could be used in an edible product such as a food product, food supplement or as a pharmaceutical composition.
- One or more embodiments involve the manufacture of food products with bioactive peptides as a consequence of the action of specific Lactococcus lactis NRRL B-50571 and/or NRRL B-50572; or bacterial preparation of the bioactive peptides, with or without other microorganisms, on a substrate within the food.
- these bioactive peptides may be separately produced and added to the food product, food supplement or pharmaceutical preparation as part of the formulation, with the purpose of reducing blood pressure, lowering LDL-cholesterol (bad cholesterol) and reducing oxidation, for better cardiovascular health.
- FIG. 3 is a typical mass spectrum corresponding to a peptide sequence collected from the WSE F1 obtained from milk fermented by L. lactis NRRL B-50571: A) Double-charged ion 362.9 m/z.; B) MS/MS Spectrum for the specified ion in A). After interpretation and comparison in database, the fragment amino acid sequence matched with ⁇ -La (f63-68);
- FIG. 4 is a diagram showing the change in blood pressure and HR during 24 h in SHR treated with milk fermented by specific L. lactis strains: (a) systolic blood pressure (SBP), (b) diastolic blood pressure (DBP) and (c) heart rate (HR).
- SBP systolic blood pressure
- DBP diastolic blood pressure
- HR heart rate
- Positive control captopril negative control saline
- whey fraction of milk fermented by L. lactis NRRL B-50572-3 35 mg protein/kg BW
- whey fraction of milk fermented by L. lactis NRRL B-50571-3 35 mg protein/kg BW
- lactis NRRL B-50572-5 50 mg protein/kg BW
- whey fraction of milk fermented by L. lactis NRRL B-50571-5 50 mg protein/kg BW
- Data is shown by means with their standard error. Each SHR group had seven animals;
- FIG. 5 is a diagram showing the change in systolic blood pressure during 4 weeks of SHR treated by L. lactis fermented milk.
- FIG. 6 is a diagram showing the diastolic blood pressure during 4 weeks of SHR treated by L. lactis fermented milk.
- FIG. 7 is a diagram showing plasma low-density lipoprotein cholesterol in SHR treated by L. lactis fermented milk for 4 weeks.
- FIG. 8 is a diagram showing plasma high-density lipoprotein cholesterol in SHR treated by L. lactis fermented milk for 4 weeks.
- FIG. 10 is a diagram showing plasma total cholesterol in SHR treated by L. lactis fermented milk for 4 weeks.
- Lactococcus lactis strains NRRL B-50571 and NRRL B-50572 have the ability to produce certain bioactive peptides having a remarkable capacity for generating an antihypertensive effect in mammals.
- These novel strains of Lactococcus lactis were deposited at the National Center for Agricultural Utilization Research, United States Department of Agriculture, 1815 N. University Street, Peoria, Ill. 61604, United States of America, in September, 2011, which are Lactococcus lactis NRRL B-50571 and NRRL B-50572. These bacteria were isolated from raw milk products and were Gram positive, catalase negative and coccal-shaped organisms.
- Lactococcus lactis was identified as Lactococcus lactis by PCR amplification of the gene acmA (Buist et al., 1995, J. Bacteriol. 177:1554-1563) with the primers PALA 4 y PALA 14 (Table 1). Strains showed the classical characteristics for Lactococcus , such as positive growth at 10 C and 4% NaCl, but lack of growth at 45 C and pH 9.6. They also presented important technological characteristics such as high proteolytic activity (8 h to coagulate litmus milk), and the ability to ferment citrate, glucose, lactose and salicin in media.
- Lactococcus lactis NRRL B-50571 or NRRL B-50572 were propagated in 10 mL of sterile lactose (5 g L ⁇ 1 ) M17 broth and incubated at 30° C. for 24 h. Fresh cultures were obtained by repeating the same procedure. Initial starter culture were prepared by allowing L. lactis strains to reach 10 6 -10 7 colony-forming units (cfu) mL ⁇ 1 as enumerated on M17 agar containing lactose (5 g L ⁇ 1 ).
- Reconstituted nonfat dry milk (10%, w/v) was sterilized at 100° C. for 20 min.
- a loop of L. lactis single pre-culture (7-8 log cfu mL ⁇ 1 ) of NRRL B-50571 or NRRL B-50572 was inoculated into sterilized milk.
- the inoculated milk was incubated for 12 h at 30° C.
- cultures were added (3% v/v) to reconstituted nonfat dry sterilized milk to get the different fermented milk batches. Incubation was carried out at 30° C. and stopped at 24 to 48 h by pasteurization at 75° C. for 1 min.
- Fermented milk was centrifuged at 20,000 ⁇ g for 10 min at 0° C. Then, supernatants were collected and ultra-filtered through 3 kDa cut-off membranes at 9,800 ⁇ g for 6 min. Permeates were collected, filtered through a 0.45 m disposable hydrophilic filter and frozen at ⁇ 80° C. until analysis were done.
- Water soluble extracts obtained after fermenting milk with L. lactis strains NRRL B-50571 or NRRL B-50572 presented high ACEI activity (>80%) and low IC 50 's ( ⁇ 25 ⁇ g/mL).
- the IC 50 is the amount of peptide content required to inhibit ACE activity by 50%.
- the ACE inhibitory activity was assayed by the method of Cushman and Cheung (Cushman and Cheung, 1971, Biochem. Phamacol. 20:1637-1648). The Cushman/Cheung method is based on the liberation of hippuric acid from hippuryl-L-histidyl-L-Leucine, catalyzed by ACE.
- Peptide profiles from WSE were obtained by RP-HPLC. Separation was carried out with a Discovery-C 18 (250 mm ⁇ 4.6 mm, 5 ⁇ m particle size, 180 ⁇ pore size) column from SUPELCO ANALYTICAL (Bellefonte, Pa., USA) with a solvent flow rate of 0.25 mL min ⁇ 1 . Once the column was equilibrated with solvent A (0.04% Trifluoroacetic acid (TFA) in water), 20 ⁇ L of the WSE were injected. Peptides were eluted with an increasing gradient of solvent B (0.03% TFA in acetonitrile) from 0% to 45% in solvent A, during 60 min.
- solvent A 0.04% Trifluoroacetic acid
- FIG. 1A shows WSE peptide fraction profiles produced by specific wild L. lactis strains monitored at 214 nm absorbance. Unfermented milk was used as a control. The area under the curve of each peptide profile was evaluated as an indirect measure of proteolysis. Results showed significant differences (P ⁇ 0.01) between fermented milk peptide profiles and the control. On the other hand, the peptide profiles obtained from milk fermented with different strains of L. lactis were similar. The first peak eluted after 12 min in all samples.
- Peptide chromatographic profiles were divided into 6 fractions and collected for further evaluation. Peptide profiles obtained at 214 nm were divided into F1-F5 fractions ( FIG. 1 A), meanwhile peptide profiles obtained at 280 nm corresponded to F6 ( FIG. 1 B). Peptide fractions F1-F6 showed remarkable IC 50 values ranging from 0.034 ⁇ 0.002 to 0.61 ⁇ 0.19 ⁇ g mL ⁇ 1 ( FIG. 2 ). Results did not show significant difference (P>0.01) between all peptide fractions IC 50 . However, the peptide fractions IC 50 values obtained from milk fermented by L.
- lactis strains NRRL B-50571 (0.076 ⁇ 0.004 and 0.034 ⁇ 0.002 ⁇ g mL ⁇ 1 for F1 and F6, respectively) and milk fermented by L. lactis NRRL B-50572 (0.041 ⁇ 0.003 and 0.084 ⁇ 0.003 ⁇ g mL ⁇ 1 for F1 and F2, respectively) showed the lowest values ( FIG. 2 ). Therefore, the results suggest that the specific wild L. lactis strains presented have remarkable ACE-Inhibitory activity. Both strains did not present a significant difference (P>0.01) in IC 50 values and proteolysis, which are related to ACE-Inhibitory activity.
- Peptide identification was performed by analyzing the different fractions by mass spectrometry using a 1100 Series LC/MSD Trap from Agilent equipped with an electro spray ionization source (LC-ESI-MS).
- the nano column was a C 18 -300 (150 mm ⁇ 0.75 ⁇ m, 3.5 ⁇ m; (AGILENT TECHNOLOGIES, INC., Palo Alto, Calif., USA).
- the sample injection volume was 1 ⁇ L.
- Solvent A was a mixture of water-acetonitrile-formic acid (10:90:0.1, v/v/v) and solvent B contained water-acetonitrile-formic acid (97:3:0.1, v/v/v).
- the gradient was based on the increment of solvent B which was initially set at 3% for 10 min and it took 23 more min to reach 65%.
- the 0.7 ⁇ L min ⁇ 1 flow rate was directed into the mass spectrometer via an electrospray interface. Nitrogen (99.99%) was used as the nebulizing and drying gas and operated with an estimated helium pressure of 5 ⁇ 10 ⁇ 3 bar.
- the needle voltage was set at 4 kV.
- Mass spectra were acquired over a range of 300-2500 mass/charge (m/z).
- the signal threshold to perform auto MS n analyses was 30,000.
- the precursor ions were isolated within a range of 4.0 m/z and fragmented with a voltage ramp from 0.35 to 1.1 V.
- Peptide sequences were obtained from mass spectrometry data using the Mascot server through the UniProtKB/Swiss-prot database sequences.
- Table 2 presents the identified sequences of peptides in the six fractions collected from milk fermented by specific L. lactis strains associated to ACEI activity.
- a typical mass spectrum of the peptide sequence DDQNPH, produced by L. lactis NRRL B-50571 fermented milk is shown in FIG. 3 .
- Samples of specific L. lactis fermented milk (prepared as previously described) for the single dose bioassay were obtained by centrifugation at 20,000 ⁇ g for 10) min at 0° C. The supernatants were collected and lyophilized with a freeze dryer until used.
- the experimental protocol was performed with forty-two male spontaneously hypertensive male rats (SHR) (4-5 weeks old, 72 ⁇ 7 g body weight (BW)) obtained from HARLAN LABORATORIES, INC, (Indianapolis, Ind., USA). SHR were weaned for eight weeks and their systolic blood pressure monitored during this period. Rats were randomly housed in pairs per cage at 21 ⁇ 2° C.
- lactis NRRL B-50572 or NRRL B-50571 were dissolved in 0.8 mL of saline. Treatments were NRRL B-50572-3 (35 mg protein/kg BW), NRRL B-50572-5 (50 mg protein/kg BW), NRRL B-50571-3 (35 mg protein/kg BW) and NRRL B-50571-5 (50 mg protein/kg BW).
- Conscious SHR received a single dose through a canula between 8:30 and 9:30 am to eliminate circadian cycles. Animals were restrained in the warming chamber for 20 min at 32° C. to detect pulsations through the caudal artery. Systolic blood pressure (SBP), diastolic blood pressure (DBP) as well as heart rate (HR) were monitored before administration and 2, 4, 6 and 24 h post-administration. Measurements were taken five times using the non-invasive blood pressure system included photoelectric sensor, amplifier, automatic inflation cuff and software (Model 229, IITC LIFE SCIENCE, Woodland Hills, Calif., USA). The animal experimental procedures were done following the guidelines and supervision of the CIAD (Centro de Investigative en Alimentación y Desarrollo), A. C. Committee of Ethics for scientific research.
- SBP stolic blood pressure
- DBP diastolic blood pressure
- HR heart rate
- SBP changes are shown in FIG. 4 a .
- Results showed the maximal SBP reductions at 6 h post oral administration.
- SHR treated with the whey fractions of milk fermented by L. lactis NRRL B-50572-5 and L. lactis NRRL B-50571-3 presented the more relevant decrement of SBP, 16.7 ⁇ 3.5 mm Hg and 17.7 ⁇ 4.0 mm Hg, respectively, although treatments were not significantly different (P ⁇ 0.05).
- the maximum decrease at 6 h was observed in animals treated with captopril which was significantly different from the treatments (P ⁇ 0.05).
- the SBP measurements 24 h post administration showed that SHR treated with the whey fraction of milk fermented by L.
- lactis NRRL B-50572-5 presented 4.3 mm Hg less than rats that were treated with captopril. These results suggest that L. lactis NRRL B-50572-5 fermented milk may have an important residual blood pressure reducing effect. Moreover, a remarkable 15.3 mm Hg SBP decrement between SHR that received the whey fraction of milk fermented by L. lactis NRRL B-50572-5 and SHR treated with saline was found. Hence, blood pressure measurements suggested an absence of dosage dependent relationship between the protein content of the whey fraction corresponding to milk fermented by L. lactis NRRL B-50571 and its ability to reduce SBP, meanwhile the whey fraction of milk fermented with L. lactis NRRL B-50572 was dosage dependent.
- FIG. 4 b shows the reduction of DBP in SHR caused by the oral administration of the whey fraction of milk fermented by specific L. lactis strains. The highest decrement of DBP was observed at 6 h post oral administration. At the same time, no significant difference was found (P ⁇ 0.05) when SHR were treated with whey fraction of milk fermented by L. lactis NRRL B-50571 at any protein content or whey fraction of fermented milk L. lactis NRRL B-50572-5. Whey fractions from milk fermented by L. lactis NRRL B-50571 as well as milk fermented with L. lactis NRRL B-50572 presented an important dosage dependent antihypertensive effect through DBP measurements. Although, captopril generated the maximum DBP reduction with each measurement, there was not a significant difference (P ⁇ 0.05) with the hypotensive effect of the whey fraction of milk fermented by L. lactis NRRL B-50572-5.
- lactis NRRL B-50571 and B-50572 was able to reduce plasma low-density lipoprotein (LDL) cholesterol by 55.4 ⁇ 3 mg/dL and 66.3 ⁇ 4 mg/dL, respectively.
- LDL low-density lipoprotein
- milk fermented by L. lactis strains may be a coadjuvant in the reduction of hypertension and hyperlipidemia and may be used as a functional food for better cardiovascular health.
- lactis strains on SHR was monitored through time. Animals were deposited in restrainers in the warming chamber for 20 min at 32° C. to detect pulsations through the caudal artery. Systolic (SBP) and diastolic (DBP) blood pressures were measured five times on each conscious animal before treatments and every week during the experiment. Measurements were obtained using the tail-cuff method between 9 and 12 h to eliminate circadian cycles.
- the non-invasive blood pressure system used in this experiment included a photoelectric sensor, an amplifier, an automatic inflation cuff and software (Model 229, IITC Life Science Inc., Woodland Hills, Calif., USA).
- SHR treated with milk fermented by L. lactis NRRL B-50571 and B-50572 presented DBP lowering-effect during the experiment ( FIG. 6 ).
- DBP the first week
- DBP measurements were not significantly different (P>0.05) between treatments.
- milk fermented by L. lactis NRRL B-50571 was able to reduce DBP by 24.5 ⁇ 6.6 mm Hg.
- captopril reduced DBP by 38.4 ⁇ 8.5 mm Hg.
- the DBP lowering-effect was not significantly different (P>0.05) between SHR treated with captopril and milk fermented by L. lactis NRRL B-50571 or B-50572.
- the most important DBP reduction 49.8 ⁇ 3.5 mm Hg was observed by the fourth week of treatment in SHR that received milk fermented by L. lactis NRRL B-50571.
- Plasma triglyceride (TG) content was also decreased by 34.7 ⁇ 3.7 mg/dL in SHR treated with L. lactis NRRL B-50572 fermented milk when compared to purified water ( FIG. 9 ). Additionally, plasma total cholesterol (TC) content was also reduced in treated SHR, although differences were not significantly different. Milk fermented by L. lactis NRRL B-50572 or B-50571 was able to reduce TC by 10 ⁇ 3.2 mg/dL and 8.6 ⁇ 2.4 mg/dL, respectively ( FIG. 10 ).
- the cholesterol lowering effect may be attributed to cholesterol removal by the L. lactis strains per se, however, this remains to be determined.
- the lowering effect on LDL-C observed in this study may also be attributed to the ingestion by SHR of dairy protein and/or peptides produced by L. lactis , including those from whey protein.
- milk fermented by specific lactic acid bacteria may be considered as a coadjuvant for the improvement of cardiovascular health.
- this is the first in vivo study that showed the antihypertensive and hypolipidemic effects of long-term consumption of fermented milk with specific L. lactis strains.
- dairy products fermented with L. lactis strains, NRRL B-50571 and NRRL B-50572 may be used as functional foods with potential benefits for cardiovascular health.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Nutrition Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Physiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Heart & Thoracic Surgery (AREA)
- Cardiology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Description
-
- L. lactis—Lactococcus lactis;
- ACE—angiotensin I-converting enzyme;
- WSE—water soluble extract;
- RP-HPLC—reverse phase high performance liquid chromatography;
- MS—mass spectrometry;
- SHR—spontaneously hypertensive rats;
- BW—body weight;
- SBP—systolic blood pressure;
- DBP—diastolic blood pressure;
- HR—heart rate;
- PP—pulse pressure;
- PWV—pulse wave velocity;
- LAB—Lactic acid bacteria;
- cfu—colony-forming units;
- LSD—Least significant difference;
- SEM—mean standard error;
- NRRL B-50571-3—milk fermented by L. lactis NRRL B-50571 (35 mg protein/kg body weight (BW);
- NRRL B-50572-3—milk fermented by L. lactis NRRL B-50572 (35 mg protein/kg BW);
- NRRL B-50571-5—milk fermented by L. lactis NRRL B-50571 (50 mg protein/kg BW);
- NRRL B-50572-5—milk fermented by L. lactis NRRL B-50572 (50 mg protein/kg BW);
- ND—not detected; and
- Vs—versus.
TABLE 1 |
Primers used for the identification |
of Lactococcus lactis strains |
Primer | Sequence | ||
PALA 4 | (5′-CTTCAACAGACAAGTCC-3′) | ||
PALA 14 | (5′-GATAAATGATTCCAAGC-3′) | ||
TABLE 2 |
Identification of peptides sequences obtained from milk fermented |
by specific wild L. lactis strains associated to ACEI activity. |
Experimental | Theoretical | Sequence | |||
Samplea | Mass | Mass | ID. | Protein fragment | Sequence |
NRRL | 723.9 | 724.3 | 1 | α-La (f63-68) | DDQNPH |
B-50571 | 1032.8 | 1033.5 | 2 | α-La (f82-89) | LDDDLTDDI |
F1 | 698.6 | 698.3 | 3 | κ-CN (f35-40) | YPSYGL |
1479.0 | 1479.7 | 4 | κ-CN (f98-110) | HPHPHLSFMAIPP | |
1035.7 | 1035.5 | 5 | α-La (f55-62) | YDTQAIVQ | |
1386.8 | 1387.7 | 6 | α-La (f100-111) | DDDLTDDIMCV | |
585.9 | 585.2 | 7 | κ-CN (f35-39) | YPSYG | |
F2 | 505.9 | 585.2 | 8 | αS1-CN (f62-66) | AESIS |
F3 | 830.1 | 830.5 | 9 | β-CN (f22-28) | SITRINK |
1051.4 | 1051.5 | 10 | αS1-CN (f80-88) | HIQKEDVPS | |
904.1 | 904.5 | 11 | κ-CN (f161-169) | TVQVTSTAV | |
F4 | 904.3 | 904.5 | 11 | κ-CN (f161-169) | TVQVTSTAV |
1038.4 | 1038.6 | 12 | αS2-CN (f115-124) | NAVPITPTLN | |
977.1 | 977.6 | 13 | β-CN (f69-77) | SLPQNIPPL | |
F5 | 1716.9 | 1717.0 | 14 | β-CN (f194-209) | QEPVLGPVRGPFPIIV |
1150.4 | 1150.7 | 15 | β-CN (f199-209) | GPVRGPFPIIV | |
977.2 | 977.6 | 13 | β-CN (f69-77) | SLPQNIPPL | |
1094.4 | 1094.6 | 16 | κ-CN (f25-33) | YIPIQYVLS | |
F6 | 904.4 | 904.5 | 11 | κ-CN (f161-169) | TVQVTSTAV |
1356.7 | 1357.7 | 17 | κ-CN (f157-169) | PEINTVQVTSTAV | |
591.8 | 592.3 | 18 | Serotransferrin (f448-453) | GYLAVA | |
NRRL | 1371.53 | 1372.7 | 19 | β-CN (f129-140) | DVENLHLPLPLL |
B-50572 | 698.6 | 698.3 | 3 | β-CN (f35-40) | YPSYGL |
F1 | 549.8 | 550.2 | 20 | β-Lg (f60-64) | ENGEC |
F2 | 904.2 | 904.5 | 11 | κ-CN (f161-169) | TVQVTSTAV |
F3 | 904.2 | 904.5 | 11 | κ-CN (f161-169) | TVQVTSTAV |
F5 | 1150.5 | 1150.7 | 15 | β-CN (f199-209) | GPVRGPFPIIV |
F6 | 922.4 | 922.4 | 21 | α-La (f86-93) | TDDIMCVK |
a= Fractions collected from milk fermented by L. lactis NRRL B-50571 and NRRL B-50572. |
Claims (8)
Priority Applications (12)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/629,398 US8865155B2 (en) | 2011-09-29 | 2012-09-27 | Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom |
PCT/IB2012/002340 WO2013046031A2 (en) | 2011-09-29 | 2012-09-28 | Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom |
MX2014003819A MX355787B (en) | 2011-09-29 | 2012-09-28 | Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom. |
US14/509,925 US9295701B2 (en) | 2011-09-29 | 2014-10-08 | Lactococcus lactis strains for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US14/525,090 US9533015B2 (en) | 2011-09-29 | 2014-10-27 | Lactococcus lactis strains |
US14/526,324 US9533016B2 (en) | 2011-09-29 | 2014-10-28 | Lactococcus lactis strains |
US15/045,709 US9717773B2 (en) | 2011-09-29 | 2016-02-17 | Lactococcus lactis strains for producing bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US15/356,328 US10092618B2 (en) | 2011-09-29 | 2016-11-18 | Composition comprising peptides made by lactococcus lactis strains |
US15/357,170 US10092619B2 (en) | 2011-09-29 | 2016-11-21 | Composition comprising peptides made by Lactococcus lactis strains |
US15/662,634 US10314884B2 (en) | 2011-09-29 | 2017-07-28 | Lactococcus lactis strains for producing bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US16/410,743 US10864247B2 (en) | 2011-09-29 | 2019-05-13 | Method for lowering low density lipoproteins |
US16/410,450 US10864246B2 (en) | 2011-09-29 | 2019-05-13 | Method for lowering triglyceride |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201161540979P | 2011-09-29 | 2011-09-29 | |
US13/629,398 US8865155B2 (en) | 2011-09-29 | 2012-09-27 | Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/509,925 Division US9295701B2 (en) | 2011-09-29 | 2014-10-08 | Lactococcus lactis strains for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
Publications (2)
Publication Number | Publication Date |
---|---|
US20140154217A1 US20140154217A1 (en) | 2014-06-05 |
US8865155B2 true US8865155B2 (en) | 2014-10-21 |
Family
ID=47996544
Family Applications (10)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/629,398 Active - Reinstated 2032-11-15 US8865155B2 (en) | 2011-09-29 | 2012-09-27 | Lactococcus lactis strains, and bacterial preparations thereof, for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects in mammals; nutritional and therapeutic products produced therefrom |
US14/509,925 Expired - Fee Related US9295701B2 (en) | 2011-09-29 | 2014-10-08 | Lactococcus lactis strains for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US14/525,090 Active US9533015B2 (en) | 2011-09-29 | 2014-10-27 | Lactococcus lactis strains |
US14/526,324 Active US9533016B2 (en) | 2011-09-29 | 2014-10-28 | Lactococcus lactis strains |
US15/045,709 Active - Reinstated US9717773B2 (en) | 2011-09-29 | 2016-02-17 | Lactococcus lactis strains for producing bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US15/356,328 Active 2033-01-02 US10092618B2 (en) | 2011-09-29 | 2016-11-18 | Composition comprising peptides made by lactococcus lactis strains |
US15/357,170 Active 2032-12-31 US10092619B2 (en) | 2011-09-29 | 2016-11-21 | Composition comprising peptides made by Lactococcus lactis strains |
US15/662,634 Active - Reinstated 2032-10-29 US10314884B2 (en) | 2011-09-29 | 2017-07-28 | Lactococcus lactis strains for producing bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US16/410,743 Active US10864247B2 (en) | 2011-09-29 | 2019-05-13 | Method for lowering low density lipoproteins |
US16/410,450 Active US10864246B2 (en) | 2011-09-29 | 2019-05-13 | Method for lowering triglyceride |
Family Applications After (9)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/509,925 Expired - Fee Related US9295701B2 (en) | 2011-09-29 | 2014-10-08 | Lactococcus lactis strains for the production of bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US14/525,090 Active US9533015B2 (en) | 2011-09-29 | 2014-10-27 | Lactococcus lactis strains |
US14/526,324 Active US9533016B2 (en) | 2011-09-29 | 2014-10-28 | Lactococcus lactis strains |
US15/045,709 Active - Reinstated US9717773B2 (en) | 2011-09-29 | 2016-02-17 | Lactococcus lactis strains for producing bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US15/356,328 Active 2033-01-02 US10092618B2 (en) | 2011-09-29 | 2016-11-18 | Composition comprising peptides made by lactococcus lactis strains |
US15/357,170 Active 2032-12-31 US10092619B2 (en) | 2011-09-29 | 2016-11-21 | Composition comprising peptides made by Lactococcus lactis strains |
US15/662,634 Active - Reinstated 2032-10-29 US10314884B2 (en) | 2011-09-29 | 2017-07-28 | Lactococcus lactis strains for producing bioactive peptides having anti-hypertensive and cholesterol-lowering effects |
US16/410,743 Active US10864247B2 (en) | 2011-09-29 | 2019-05-13 | Method for lowering low density lipoproteins |
US16/410,450 Active US10864246B2 (en) | 2011-09-29 | 2019-05-13 | Method for lowering triglyceride |
Country Status (3)
Country | Link |
---|---|
US (10) | US8865155B2 (en) |
MX (1) | MX355787B (en) |
WO (1) | WO2013046031A2 (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3590519A4 (en) * | 2017-03-03 | 2020-12-16 | Morinaga Milk Industry Co., Ltd. | Glp-1 secretagogue and composition |
CN107771940A (en) * | 2017-10-17 | 2018-03-09 | 陕西科技大学 | A kind of preparation method of oxidation resistant Lactobacillus casei fermentation sheep breast |
KR102224707B1 (en) * | 2019-05-24 | 2021-03-08 | 부산대학교 산학협력단 | Composition for preventing or treating hyperlipidemia comprising bioactive peptides |
EP3922309A1 (en) | 2020-06-12 | 2021-12-15 | Innovación en Gestón y Conservación de Ungulados S.L. (Ingulados) | Novel lactococcus lactis strain for the production of bioactive compounds having antimicrobial effect |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050031602A1 (en) | 2003-04-07 | 2005-02-10 | Chr. Hansen A/S | Composition with heart rate reducing properties |
WO2007096855A2 (en) * | 2006-02-24 | 2007-08-30 | Teagasc, The Agriculture And Food Development Authority | Angiotensin-i-converting enzyme inhibitory |
US20070203060A1 (en) * | 2004-03-01 | 2007-08-30 | Zvi Sidelman | Casein Derived Peptides And Therapeutic Uses Thereof |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH09188694A (en) * | 1996-01-10 | 1997-07-22 | Otsuka Shokuhin Kk | Peptide, angiotensinase inhibiting composition and their production |
-
2012
- 2012-09-27 US US13/629,398 patent/US8865155B2/en active Active - Reinstated
- 2012-09-28 MX MX2014003819A patent/MX355787B/en active IP Right Grant
- 2012-09-28 WO PCT/IB2012/002340 patent/WO2013046031A2/en active Application Filing
-
2014
- 2014-10-08 US US14/509,925 patent/US9295701B2/en not_active Expired - Fee Related
- 2014-10-27 US US14/525,090 patent/US9533015B2/en active Active
- 2014-10-28 US US14/526,324 patent/US9533016B2/en active Active
-
2016
- 2016-02-17 US US15/045,709 patent/US9717773B2/en active Active - Reinstated
- 2016-11-18 US US15/356,328 patent/US10092618B2/en active Active
- 2016-11-21 US US15/357,170 patent/US10092619B2/en active Active
-
2017
- 2017-07-28 US US15/662,634 patent/US10314884B2/en active Active - Reinstated
-
2019
- 2019-05-13 US US16/410,743 patent/US10864247B2/en active Active
- 2019-05-13 US US16/410,450 patent/US10864246B2/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050031602A1 (en) | 2003-04-07 | 2005-02-10 | Chr. Hansen A/S | Composition with heart rate reducing properties |
US20070203060A1 (en) * | 2004-03-01 | 2007-08-30 | Zvi Sidelman | Casein Derived Peptides And Therapeutic Uses Thereof |
WO2007096855A2 (en) * | 2006-02-24 | 2007-08-30 | Teagasc, The Agriculture And Food Development Authority | Angiotensin-i-converting enzyme inhibitory |
Non-Patent Citations (7)
Title |
---|
Fugslang, et al., "Cardiovascular Effects of Fermented Milk Containing Angiotensin-Converting Enzyme Inhibitors Evaluated in Permanently Catheterized, Spontaneously Hypertensive Rats", Applied and Environmental Microbiology 68(7), Jul. 2002, pp. 3566-3569. |
Gutierrez-Mendez, et al., "Phenotypic and genotypic characteristics of Lactococcus lactis strains isolated from different ecosystems", Canadian Journal of Microbiology (56), May 2010, pp. 432-439. |
Rodriguez-Figueroa, et al., "Angiotensin-converting enzyme inhibitory activity of milk fermented by wild and industrial Lactococcus lactis strains", Journal of Dairy Sicence; 93(11), Nov. 2010, pp. 5032-5038. |
Rodríguez-Figueroa, J. C., González-Córdova, A. F., Astiazaran-García, H. and Vallejo-Cordoba, B. 2013. Hypotensive and heart rate lowering effects in rats receiving milk fermented by specific Lactococcus lactis strains. ISSN: 0007-1145.British J. Nutr., 109, 827-833. |
Rodríguez-Figueroa, J. C., González-Córdova, A. F., Astiazaran-García, H., Hernández-Mendoza, A. and Vallejo-Cordoba, B. 2013. Antihypertensive and hypolipidemic effect of milk fermented by specific Lactococcus lactis strains. ISSN: 0022-0302. Journal of Dairy Science. 96(7): 4094-4099. |
Rodríguez-Figueroa, J. C., González-Córdova, A. F., Torres-Llanez, M. J., Garcia, H. S. and Vallejo-Cordoba, B. 2012. Novel angiotensin I-converting enzyme inhibitory peptide produced in fermented milk by specific wild Lactococcus lactis strains. ISSN 0022-0302. Journal of Dairy Science. 95(10): 5536-5543. |
Torres-Llanez, et al., "Angiotensin-converting enzyme inhibitory activity in Mexican Fresco cheese", Journal of Dairy Science 94(8), Jul. 2011, pp. 3974-3800. |
Also Published As
Publication number | Publication date |
---|---|
MX2014003819A (en) | 2016-03-04 |
US20150050255A1 (en) | 2015-02-19 |
US20140154217A1 (en) | 2014-06-05 |
US9295701B2 (en) | 2016-03-29 |
WO2013046031A2 (en) | 2013-04-04 |
US20150051159A1 (en) | 2015-02-19 |
US10864247B2 (en) | 2020-12-15 |
US20190298793A1 (en) | 2019-10-03 |
US20150064271A1 (en) | 2015-03-05 |
US10314884B2 (en) | 2019-06-11 |
US10092619B2 (en) | 2018-10-09 |
WO2013046031A3 (en) | 2013-07-25 |
US9533015B2 (en) | 2017-01-03 |
US20170065666A1 (en) | 2017-03-09 |
MX355787B (en) | 2018-04-27 |
US20170065665A1 (en) | 2017-03-09 |
US20160158296A1 (en) | 2016-06-09 |
US10864246B2 (en) | 2020-12-15 |
US20190269751A1 (en) | 2019-09-05 |
US20170326198A1 (en) | 2017-11-16 |
US9533016B2 (en) | 2017-01-03 |
US10092618B2 (en) | 2018-10-09 |
US9717773B2 (en) | 2017-08-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10864247B2 (en) | Method for lowering low density lipoproteins | |
Beltrán-Barrientos et al. | Invited review: Fermented milk as antihypertensive functional food | |
Takano | Anti-hypertensive activity of fermented dairy products containing biogenic peptides | |
Minervini et al. | Fermented goats’ milk produced with selected multiple starters as a potentially functional food | |
RU2276689C2 (en) | Method for preparing hypotensive peptide-containing foodstuff, foodstuff prepared by this method and foodstuff | |
JP2011504363A (en) | Production of improved bioactive peptides | |
KR20090122454A (en) | Agent for reducing visceral fat | |
KR101553358B1 (en) | Anti-allergic agent | |
US20100021444A1 (en) | Blood-adiponectin-concentration increase-accelerator and/or decrease-inhibitor thereof and visceral fat accumulation inhibitor | |
Nakamura | Studies on anti-hypertensive peptides in milk fermented with Lactobacillus helveticus | |
EP1640447A1 (en) | BIOACTIVE PEPTIDE-PRODUCING STRAINS OF i ENTEROCOCCUS FAECALIS /i , BIOACTIVE PEPTIDES AND APPLICATIONS THEREOF | |
US7785633B2 (en) | Agent for preventing or suppressing hepatopathy and functional food for preventing or suppressing hepatopathy | |
US20230180778A1 (en) | Milk fermentation process | |
WO2018034279A1 (en) | Composition for controlling acquired immune function suppression due to anti-influenza drug, and production method for same | |
Acharya | Production of functional food, Fruit ravioli with antihypertensive peptides | |
Hayakawa | Beneficial effects of fermented milk |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: GALLAND, BELINDA VALLEJO, MEXICO Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:RODRIGUEZ FIGUEROA, JOSE CARLOS;REEL/FRAME:030851/0202 Effective date: 20130220 Owner name: CENTRO DE INVESTIGACION EN ALIMENTACION Y DESARROL Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:RODRIGUEZ FIGUEROA, JOSE CARLOS;REEL/FRAME:030851/0202 Effective date: 20130220 Owner name: GONZALEZ CORDOVA, AARON FERNANDO, MEXICO Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:RODRIGUEZ FIGUEROA, JOSE CARLOS;REEL/FRAME:030851/0202 Effective date: 20130220 |
|
STCF | Information on status: patent grant |
Free format text: PATENTED CASE |
|
MAFP | Maintenance fee payment |
Free format text: PAYMENT OF MAINTENANCE FEE, 4TH YR, SMALL ENTITY (ORIGINAL EVENT CODE: M2551) Year of fee payment: 4 |
|
FEPP | Fee payment procedure |
Free format text: MAINTENANCE FEE REMINDER MAILED (ORIGINAL EVENT CODE: REM.); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY |
|
LAPS | Lapse for failure to pay maintenance fees |
Free format text: PATENT EXPIRED FOR FAILURE TO PAY MAINTENANCE FEES (ORIGINAL EVENT CODE: EXP.); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY |
|
STCH | Information on status: patent discontinuation |
Free format text: PATENT EXPIRED DUE TO NONPAYMENT OF MAINTENANCE FEES UNDER 37 CFR 1.362 |
|
FP | Lapsed due to failure to pay maintenance fee |
Effective date: 20221021 |
|
PRDP | Patent reinstated due to the acceptance of a late maintenance fee |
Effective date: 20230209 |
|
FEPP | Fee payment procedure |
Free format text: PETITION RELATED TO MAINTENANCE FEES FILED (ORIGINAL EVENT CODE: PMFP); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY Free format text: PETITION RELATED TO MAINTENANCE FEES GRANTED (ORIGINAL EVENT CODE: PMFG); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY Free format text: SURCHARGE, PETITION TO ACCEPT PYMT AFTER EXP, UNINTENTIONAL. (ORIGINAL EVENT CODE: M2558); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY |
|
MAFP | Maintenance fee payment |
Free format text: PAYMENT OF MAINTENANCE FEE, 8TH YR, SMALL ENTITY (ORIGINAL EVENT CODE: M2552); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY Year of fee payment: 8 |
|
STCF | Information on status: patent grant |
Free format text: PATENTED CASE |